Prevalence and Molecular Characterization of ESBL/pAmpC Producing Faecal Escherichia coli Strains with Widespread Detection of CTX-M-15 Isolated from Healthy Poultry Flocks in Eastern Algeria.

The intensification of livestock farming has led to the widespread use of massive amounts of antibiotics worldwide. Poultry production, including white meat, eggs and the use of their excreta as fertiliser, has been identified as one of the most crucial reservoirs for the emergence and spread of resistant bacteria, including E. coli in poultry as an important opportunistic pathogen representing the greatest biological hazard to human and wildlife health. Thus, this study aimed to analyse E. coli in the faecal carriage of healthy poultry flocks and to investigate the phenotypic and genotypic characteristics of antimicrobial resistance, including integrons genes and phylogenetic groups. A total of 431 cloacal swabs from apparently healthy poultry from four regions in Eastern Algeria from December 2021 to October 2022. 360 E. coli were isolated; from broilers (n=151), broiler breeders (n=91), laying hens (n=72), and breeding hens (n=46). Among this, 281 isolates exhibited multidrug resistance (MDR) phenotype, 17 of the 360 E. coli isolates exhibited ESBL, and one isolate exhibited both ESBL/pAmpC. A representative collection of 183 among 281 MDR E. coli was selected for further analysis by PCR to detect genes encoding resistance to different antibiotics, and sequencing was performed on all positive PCR products of blaCTX-M and blaCMY-2 genes. Phylogenetic groups were determined in 80 E. coli isolates (20 from each of the four kinds of poultry). The blaCTX-M gene was found in 16 (94.11 %) ESBL-producing E. coli isolates within 11 strains co-expressing the blaSHV gene and 8 strains co-expressing the blaTEM gene. Sequence analysis showed frequent diversity in CTX-M-group-1, with blaCTX-M-15 being the most predominant (n=11), followed by blaCTX-M-1 (n=5). The blaCMY-2 gene was detected only in one ESBL/pAmpC isolate. Among the 183 tested isolates, various antimicrobial resistance genes were found (number of strains) blaTEM (n=121), blaSHV (n=12), tetA (n=100), tetB (n=29), sul1(n=67), sul2 (n=32), qnrS (n=45), qnrB (n=10), qnrA (n=1), catA1(n=13), aac-(6')-Ib (n=3). Furthermore, class 1 and class 2 integrons were found in 113 and 2 E. coli, respectively. The isolates were classified into multiple phylogroups, including A (35%), B1 (27.5%), B2 and D each (18.75%). The detection of integrons and different classes of resistance genes in the faecal carriage of healthy poultry production indicates that commensal E. coli could potentially act as a reservoir for antimicrobial resistance, posing a significant One Health challenge encompassing the interconnected domains of human, animal health and the environment. Here, we present the first investigation to describe the diversity of blaCTX-M producing E. coli isolates with widespread detection of CTX-M-15 and CTX-M-1 in healthy breeders (Broiler and breeding hens) in Eastern Algeria.

Exploring Sorbus torminalis Leaves: Unveiling a Promising Natural Resource for Diverse Chemical and Biological Applications.

Sorbus torminalis (L.) Crantz has a rich history of versatile applications spanning the fields of medicine and nutrition. It is noteworthy that the decoction obtained from S. torminalis leaves is a traditional treatment method against both diabetes and stomach disorders. Phytochemical profiling determined by HPLC/MS-MS. The effects of the extracts on cell viability were investigated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) method against MDA-MB-231 cell line (human breast adenocarcinoma).The ethanol/water extract contained more concentration of total phenolic (91.41 mg gallic acid (GAE) equivalent /gr) and flavanoid (29.10 mg rutin (RE) equivalent/gr) in the tested extract (p<0.05). Resulting of HPLC analysis, the chemical constituents varied depending on the solvents and chlorogenic acid, hyperoside, isoquercetin, delphindin-3,5-diglucoside, procyanidin B2, epicatechin, neochlorogenic acid, 3,5-dicaffeoylquinic acid were identified in all extracts. Overall, ethanol, n-hexane and ethyl acetate extracts showed the highest inhibition for the tyrosinase enzyme. The effect of leaf extracts of S. torminalis on antimicrobial, biofilm inhibitory, and anticancer activities was examined. Based on outcomes of our study recognize this plant as a critical source of medically active chemicals for feasible phytopharmaceutical and nutraceutical applications, providing the first scientific insight into the detailed biological and chemical profiles of S. torminalis.

Investigation of Toxoplasma gondii Seropositivity in Pregnant Women in Kastamonu Province, Turkey

Objective: Toxoplasma gondii (T. gondii) is a protozoan parasite that infects most warm-blooded animal species and causes toxoplasmosis. Especially infections that occur during pregnancy can lead to serious clinical symptoms. This study retrospectively revealed the T. gondii seroprevalence of pregnant women in Kastamonu province, Turkey. Methods: Anti-T. gondii IgM and IgG positivity of 1.294 pregnant women between the ages of 15-44 years who applied to the Obstetrics and Gynecology Outpatients of Kastamonu Training and Research Hospital from January 2018 to January 2022 were investigated retrospectively. The IgG avidity test was performed for both anti-T. gondii IgM and IgG positivity. Results: Anti-T. gondii IgM and IgG seropositivity were determined as 1.1% (n=14) and 20.3% (n=263), respectively. Anti-T. gondii IgM and IgG positivity were detected together in 11 pregnant women. IgG avidity test results of only six pregnant women could be reached, two pregnant had high IgG avidity, and four pregnant had low IgG avidity. Anti-T. gondii IgG positivity rate increased with increasing age (p=0.039). Conclusion: The number of seronegative pregnant women was considered high in Kastamonu. It is significant for expectant mothers to know about prevention methods in order not to acquire toxoplasmosis.

Presence of quorum sensing system, virulence genes, biofilm formation and relationship among them and class 1 integron in carbapenem-resistant clinical Pseudomonas aeruginosa isolates.

Carbapenems are the most effective agents for treating clinical P. aeruginosa (PsA) infections. During an infection, a quorum-sensing (QS) system and its regulating virulence genes have a great role. The aim of the study was to detect the presence of a las and rhl QS system and related virulence genes, biofilm formation and a class 1 (Cls1) integron. A total of 52 carbapenem-resistant PsA (CRPsA) isolates obtained from Kastamonu, Turkey was analyzed. For the isolation and identification of CRPsA isolates, a conventional culture method, an automated VITEK-2 compact system, and oprL gene-based molecular technique were applied. The two QS system genes were detected in 51 (98.1%), and co-existed of four two QS system genes (lasI/R and rhIl/R genes) were determined in 41 (78.8%) of the isolates. algD, lasB, toxA and aprA genes were detected in between 46.1 and 88.5%, and co-existence of four two QS system genes with four virulence genes were detected in 40.4% of the isolates. Biofilm formation using microtiter plate assay and slime production using Congo Red Agar and Cls1 integron were determined in 84.6%, 67.3% and 51.9% of the isolates, respectively. According to statistical analyses results, there was a significant positive correlation (p < .10) between the las and the rhl systems and a strongly and positive correlation (p < .01 or p < .05) between the rhl system-three virulence genes and slime production-and among some virulence genes. In conclusion, the CRPsA isolates tested in the study are highly virulent and QS systems have a significant role in pathogenesis.

Isolation and Characterization of Lytic Bacteriophages from Wastewater with Phage Therapy Potentials Against Gram-Negative Bacteria.

OBJECTIVE: The increase of multidrug resistance in bacteria has increased the efforts in search of alternative methods. The aim of the present study was to isolate and characterize the lytic phages and assess their lytic activity against a number of gram-negative bacteria. MATERIALS AND METHODS: The phages and their respective hosts were isolated from wastewater collected from the municipal sewer system of Trabzon, Turkey. The lytic activities of phage were determined using the agar spot test. The identification and antibiotic susceptibility of host bacteria were determined using matrix-assisted laser desorption ionization time-of-flight mass spectrometry and Phoenix 100, respectively. The phages were characterized morphologically using transmission electron microscopy. One of the phages, Enteroc21, which has a broad-host-range, was further characterized by genome restriction endonuclease analysis and burst size. RESULTS: Two phages infected strains of four different species, nine phages were able to infect 2-4 strains belonging to one or two species, and three phages showed lytic activity against only the hosts from which they were isolated. All phages belonged to the Siphoviridae, Myoviridae, and Podoviridae family based on trans- mission electron microscopy morphology. The Enteroc21 had more than 100 kb genome size and a burst size of 180 per infected cell. Most of the host strains were resistant to ampicillin, amoxicillin-clavulanic acid, and in particular, Achromobacter xylosoxidans TRAX 13 was multidrug-resistant showing resistance to cefepime, aztreonam, gentamicin, netilmicin, and ciprofloxacin. CONCLUSION: This study showed that the isolated phages have the potential to be used in phage therapy against various bacterial infections, including multidrug-resistant bacteria.

Identification and antifungal susceptibility of Saprochaete clavata from invasive infections in Turkey.

Saprochaete clavata is an emerging opportunistic pathogen, that causes life-threatening infections, but there are limited evidence and information about the evaluation of in vitro antifungal susceptibility test results. The aim of this study was to determine S. clavata isolates from clinical specimens and to investigate their in vitro antifungal susceptibility. S. clavata was identified by API ID20C AUX (BioMérieux, Brussels, Belgium), MALDI TOF (Bruker Daltonik, Germany), and ITS gene region sequencing. In vitro susceptibility tests were performed using Sensititre YeastOne (TREK Diagnostic System, East Grinstead, UK). During the study period, 4,736 fungi were isolated from various clinical samples and, S. clavata was identified in eight patients with underlying diseases namely, pancreatic neoplasma, acute myeloid leukaemie, follicular lymphoma, cholelithiasis. Anidulafungin and micafungin minimum inhibitory concentration values were 1-2 and 1-4 mg/L, respectively, while those of the azole group antifungals were much lower. This is the first study in Turkey reporting isolation, identification and antifungal susceptibilities of S. clavata from clinical specimens. Higher MIC values seen in some isolates suggest that continuous monitoring of sensitivity rates and observation of regional differences will thus be useful guides in determining infection control and antifungal use policies.

Chemical characterization, computational analysis and biological views on Daphne gnidioides Jaub. & Spach extracts: Can a new raw material be provided for biopharmaceutical applications?

The scientific world tends to turn to natural products such as medicinal and aromatic plants because of the inadequacy of commercially available synthetic drugs as antibiotics or anticancer, and their adverse effects on healthy tissues. One of these plants is Daphne gnidioides Jaub. & Spach, which belongs to the Thymelaeaceae family, and there is no data in the literature on its biological activity. This study is aimed to elucidate the chemical profiles and in vitro anticancer, antibacterial and DNA protection and enzyme inhibitory properties of methanol extracts of root, stem, and leaf of D. gnidioides Jaub. & Spach. Polyphenolic components of the extracts were characterized by HPLC-MS/MS. The highest phenolic content was detected in the leaf extract (TIPC = 43.5 ± 0.5 mg/g DE), followed by stem (TIPC = 27.3 ± 0.7 mg/g DE) and root (TIPC = 18.3 ± 0.2 mg/g DE) extracts. Vicenin-2 and 3-O-p-coumaroyl-5-O-caffeoylquinic acid were the main identified compounds in leaf and both root and stem extracts, respectively. The extracts did not show any protective effect on DNA against experimental Fenton's reagent. The minimum inhibitory concentration and the minimum bactericidal concentration values for the root and leaf extracts against tested bacterial strains ranged from 31.25 to 500 µg/mL. After 48 h interaction of the cancer cell lines with the extracts, only the stem extract had significant cytotoxicity on HeLa cells (IC50 = 86.16 µg/mL). No remarkable activity of the extracts, which was tested against MDA-MB-231, was detected (IC50 > 1000 µg/mL). These data showed that D. gnidioides Jaub. & Spach stem extract inhibited the survival of HeLa cells in a time-dependent manner. After the treatment of IC50 concentration of stem extract with HeLa cells, an increase in LC3-II autophagic gene expression was detected. Also, the extracts exhibited significant tyrosinase inhibitory effects which were confirmed by molecular docking. To sum up, the tested extracts could be used as a starting point for the development of new multifunctional drugs.